鼻咽癌细胞株SUNE1中EB病毒LMP1全基因克隆及部分？…

目的 研究广东地区鼻咽癌（ＮＰＣ）细胞株ＳＵＮＥ１中ＥＢ病毒ＬＭＰ１基因的结构特征。方法 利用聚合酶链反应从ＳＵＮＥ１细胞基因组中扩增ＬＭＰ１全长ＤＮＡ，并克隆到ｐｃＤＮＡ３载体中，采用Ｓａｎｇｅｒ双脱氧末端终止法测定ＳＵＮＥ－ＬＭＰ１ ３个外显子及２个内含子覆盖的序列，并与病毒原型Ｂ９５－８－ＬＭＰ１进行比较分析。结果 克隆到ｐｃＤＮＡ３中的ＳＵＮＥ１－ＬＭＰ１全基因长度为２．６ｋｂ，测序长度为

DNA cloning and partial sequence analysis of EBV LMP1 gene isolated from human nasopharyngeal carcinoma cell line SUNE1]

OBJECTIVE: To study the variation of EBV LMP1 encoding gene isolated from human nasopharyngeal carcinoma cell line SUNE1 in Guangdong. METHODS: The EBV LMP1 gene was amplified from the SUNE1 cell genome by PCR and then the recombinant vector was constructed by inserting the PCR fragment into pcDNA3. The encoding EBV LMP1 spanning from exon1 to exon3 in recombinant vector was sequenced by the Sequence Analyzer. RESULTS: The 1,312 bp encoding EBV LMP1 pieces in 2.6 kb PCR fragments were compared with the same EBV segments in B95-8 cell line. The data indicated that the rate of nucleotide sequence homology between the two fragments is 98.5% and the rate of amino acid is 96%. The restricted enzyme site of Xho I in exon1 was deleted in SUNE1 but the 30 bp deletion at the carboxyl terminus in most Chinese NPC LMP1 gene was not present. CONCLUSION: Although the LMP1 gene derived from SUNE1 had greater tumorigenicity than that derived from B95-8 cell, the high homology rate of nucleotide and amino acid sequences between them indicated that it was not the result from the variation of certain nucleotide sites but the change in amino acid domain.